acetone fixation of frozen sections

[MSturm]

We are searching for a protocol for acetone fixation for frozen sections.  The problem with the fixation that we are currently applying is that there is nuclear smearing and loss of cellular detail.  Routinely we fix for 10 minutes in cold acetone IN the freezer, then wash with 2 changes of buffer at 5 minutes each.  Anyone have some comments for us?

thanks
Mary Sturm

[Cardio]

We are searching for a protocol for acetone fixation for frozen sections.  The problem with the fixation that we are currently applying is that there is nuclear smearing and loss of cellular detail.  Routinely we fix for 10 minutes in cold acetone IN the freezer, then wash with 2 changes of buffer at 5 minutes each.  Anyone have some comments for us?

thanks
Mary Sturm

You can try to increase the time to 20 minutes and use fresh acetone or you can try methanol. IF you start out with autolysis tissue then you can get nuclear smearing. What is the tissue?

[MT Scientist]

I cut cryosections and immediately fix in 95% ethanol prior to air-drying.

See http://www.pathologyinnovations.com/frozen_section_technique.htm

[Dunia]

Do you wash the sections with buffer immediately after taking them out of a jar with acetone? This could be the problem.
Try to air-dry the slides (about 20 min RT) before proceeding with aqueous buffers.